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Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR
Authors:Jesús Mercado-Blanco  Dolores Rodríguez-Jurado  Encarnación Pérez-Artés  Rafael M Jiménez-Díaz
Institution:(1) Departamento de Protección de Cultivos, Instituto de Agricultura Sostenible (IAS), Consejo Superior de Investigaciones Científicas (CSIC), Apartado 4084, 14080 Córdoba, Spain
Abstract:Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.
Keywords:molecular diagnosis  Olea europaea  pathogen-free certification programs  Verticillium wilt
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